Isolation and characterization of a photoaffinity-labeled peptide from the catalytic site of prenyltransferase.
Isolation and characterization of a photoaffinity-labeled peptide from the catalytic site of prenyltransferase.
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从异戊二烯基转移酶的催化位点分离和表征光亲和标记的肽。
DOI：
10.1021/bi00516a007
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发表时间：
1981
期刊：
影响因子：
2.9
通讯作者：
Rilling,HC
中科院分区：
文献类型：
--
作者：
Brems,DN;Bruenger,E;Rilling,HC
材料和方法酶制剂。改变了Reed&Ring(1975)的纯化程序，加入了CaCl2沉淀步骤，以降低脂质含量。在原程序下，通常可获得的鸡肝脂肪含量较高，导致第一次离心法和第一次硫酸铵沉淀法分离效果不佳。一般情况下，4公斤肝脏在9.4 L 50 mM咪唑缓冲液中解冻，pH 7.0，含有10 mM 2-巯基乙醇，在瓦林搅拌器中匀浆1min。
Materials and MethodsEnzyme Preparation. The purification procedure of Reed & Rilling (1975) was altered by including a CaCl2 precipitation step in order to reduce the lipid content. Under the original procedure, the high lipid content of chicken liver, usually available, resulted in poor separations in the first centrifugation and the first ammonioum sulfate precipitation. Typically, 4 kg of liver was thawed in 9.4 L of 50 mM imidazole buffer, pH 7.0, containing 10 mM 2-mercaptoethanol and homogenized in a Waring blender for 1 min.